Human Growth Hormone

Growth hormone (somatotropin) size 191 amino

acids mol wt. 22125 kd is secreted by Anterior
Pituitary

Growth hormone stimulates production of

insulin-like growth factor 1.

Insulin-like growth factor 1 is an essential

component of the promotion of growth in
children, and in adults

It controls metabolism
Infants and children who lack sufficient endogenous levels
of human growth hormone
Patients with chronic renal insufficiency (defective
kidneys),
Turner syndrome
Respond to treatment with growth hormone

It stimulates
Tissue and bone growth,
increases protein synthesis
Mineral retention
Decreases body fat storage
In 1982, human insulin became the first
pharmaceuticals produced by recombinant DNA
technologies for commercial use.

Since then, several other human proteins with

medicinal value have been synthesized in bacteria.
Some of the first human proteins to be produced in
microorganisms were :

• Blood-clotting factor VIII (lacking in individuals

with one type of hemophilia)
• Plasminogen activator (a protein that disperses
blood clots)
• Human growth hormone (a protein deficient in
certain types of dwarfism).
In 1985, hGH became the second genetically
engineered pharmaceutical approved for use in
humans by the U.S. Food and Drug Administration.

The first recombinant growth hormone was called

somatrem (Protropin)

It had an amino acid sequence that was identical to

that of human growth hormone, except that there was
an extra methionine residue at the N-terminal end of
the peptide chain
Only growth hormones from humans or from closely
related primates will function in humans.

Thus, prior to 1985, the major source of growth

hormone suitable for treatment of humans was from
human cadavers

hGH, which is required for normal growth, is a single

polypeptide chain 191 amino acids in length
To obtain expression in E. coli, the hGH coding
sequence must be placed under the control of E.
coli regulatory elements.

Therefore, the hGH coding sequence was joined to

the promoter and ribosome-binding sequences of
the E. coli lac operon
To accomplish this, a HaeIII cleavage site in the
nucleotide-pair triplet specifying codon 24 of hGH
was used to fuse a synthetic DNA sequence encoding
amino acids 1–23 to a partial cDNA sequence
encoding amino acids 24–191.

This unit was then inserted into a plasmid carrying

the lac regulatory signals and introduced into E. coli
by transformation
The hGH produced in E. coli in these first experiments
contained methionine at the amino terminus (the
methionine specified by the ATG initiator codon).

Native hGH has an amino-terminal phenylalanine: a

methionine is initially present but is then enzymatically
removed.

E. coli also removes many amino-terminal methionine

residues posttranslationally. However, the excision of the
terminal methionine is sequence-dependent, and E. coli
cells do not excise the amino-terminal methionine residue
from hGH.

Nevertheless, the hGH synthesized in E. coli was found to

be fully active in humans despite the presence of the extra
amino acid.
More recently, a DNA sequence encoding a signal peptide
(the amino acid sequence required for transport of
proteins across membranes) has been added to an hGH
gene construct.

With the signal sequence added, hGH is both secreted

and correctly processed The methionine residue is
removed with the rest of the signal peptide during the
transport of the primary translation product across the
membrane.

This product is identical to native hGH.

in 2004, the U.S. Food and Drug Administration (FDA)
approved the use of recombinant human growth
hormone for individuals whose short stature was caused
by a variety of medical conditions other than human
growth hormone deficiency.
Native human growth hormone binds to both growth hormone
and prolactin receptors that occur on a number of different cell
types.

To avoid unwanted side effects during therapy, it is desirable

that human growth hormone bind only to growth hormone
receptors.

Site-specific mutagenesis of the cloned human growth

hormone cDNA was used to change some of the amino acid
side chains that act as ligands for Zn (i.e., His-18, His-21, and
2+

Glu-174), because the ion is required for the high-affinity

binding of human growth hormone to the prolactin Receptor

These modifications yielded human growth hormone

derivatives that bound to the growth hormone receptor but not
to the prolactin receptor.
Short half-life in plasma, human growth hormone therapy
currently requires subcutaneous injection once a day.

This treatment is both inconvenient and expensive.

The extracellular domain of the human growth hormone

receptor was fused to human growth hormone

This construct has a very strong tendency to dimerize as the

growth hormone moiety from one molecule binds with the
receptor portion of another molecule.

When this growth hormone construct was tested in rats, a

single injection promoted growth for 10 days
Another method that has been devised to prolong the
active lifetime of human growth hormone includes fusing
the coding sequences for the C-terminal end of human
growth hormone with the N-terminal end of human serum
albumin. This fusion protein is called Albutropin

The stabilization of the human growth hormone portion of

Albutropin reflects the stability of human serum albumin,
which has a half-life in serum of about 19 days.